Development and evaluation of a human T-cell leukemia virus type I serologic confirmatory assay incorporating a recombinant envelope polypeptide.
A recombinant protein derived from the gp21 region of the human T-cell leukemia virus type I (HTLV-I) env gene was synthesized in Escherichia coli and purified by reversed-phase high-performance liquid chromatography. The purified protein was free of contaminating bacterial proteins and retained reactivity with human HTLV-I- and HTLV-II-positive sera and a gp21 monoclonal antibody. An immunoblot procedure using the recombinant polypeptide in conjunction with native viral proteins was more sensitive than the conventional immunoblot and radioimmunoprecipitation confirmatory assays for detection of antibodies to HTLV-I and HTLV-II env-encoded gene products. The recombinant protein was equally reactive with sera from polymerase chain reaction-confirmed HTLV-I or HTLV-II infections. Furthermore, on the basis of the differential reactivities of gp21-positive sera with the HTLV-I p19 and p24 gag-encoded proteins, an algorithm was proposed to distinguish exposure to HTLV-I from exposure to HTLV-II. These results establish the utility of a modified immunoblot assay incorporating a recombinant envelope polypeptide as an alternative to existing HTLV-I-confirmatory assays.
Lillehoj
E P
EP
Cambridge Biotech Corp., Rockville, Maryland 20850.
Alexander
S S
SS
Dubrule
C J
CJ
Wiktor
S
S
Adams
R
R
Tai
C C
CC
Manns
A
A
Blattner
W A
WA
D000914:Antibodies, Viral / Q000032:analysis*; D005069:Evaluation Studies as Topic; D015686:Gene Products, env; D015490:HTLV-I Infections / Q000175:diagnosis / Q000276:immunology; D015491:HTLV-II Infections / Q000175:diagnosis; D015368:Human T-lymphotropic virus 1 / Q000276:immunology*; D015367:Human T-lymphotropic virus 2 / Q000276:immunology; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D015531:Radioimmunoprecipitation Assay; D011994:Recombinant Proteins / Q000276:immunology; D015682:Retroviridae Proteins, Oncogenic / Q000276:immunology; D054299:env Gene Products, Human Immunodeficiency Virus
D000914:Antibodies, Viral; D015686:Gene Products, env; D011994:Recombinant Proteins; D015682:Retroviridae Proteins, Oncogenic; D054299:env Gene Products, Human Immunodeficiency Virus; C055265:gp21 protein, Human T-lymphotropic virus 1